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Vicia sativa ILP molecular marker primers and application thereof in variety identification

A technology of arrowhead pea and molecular markers, applied in the biological field, can solve the problems of lack of whole genome data, achieve high sensitivity and accuracy, convenient operation, and low cost

Active Publication Date: 2019-11-01
LANZHOU UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the lack of whole-genome data, related studies on the arrow tube pea have not been reported yet.

Method used

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  • Vicia sativa ILP molecular marker primers and application thereof in variety identification
  • Vicia sativa ILP molecular marker primers and application thereof in variety identification
  • Vicia sativa ILP molecular marker primers and application thereof in variety identification

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0063] 1. Identification material DNA extraction

[0064] Referring to the SDS method, the specific operation steps are as follows:

[0065] 1) Take 5 g of collected fresh leaves, put them into a mortar and grind them into powder, add 600 μl of DNA buffer (30% SDS lysate) into the mortar, and mix well.

[0066] 2) Pour the homogeneous solution into a centrifuge tube and place in a water bath at 60°C for 10 minutes, and shake it every 3-4 minutes during the process.

[0067] 3) Take it out and cool to room temperature, add 195 μl of 5M KAc solution, mix well and place on ice for 5 minutes.

[0068] 4) Add 600 μl of chloroform isoamyl alcohol (24:1) (operate in a fume hood with the ventilation system turned on), and mix well.

[0069] 5) Let it cool down to room temperature, centrifuge in a centrifuge at 12000r / min for 10min, and pipette 360μl of the supernatant into a clean centrifuge tube.

[0070] 6) Add 240 μl isopropanol and 36 μl 3M NaAc solution to the supernatant, mix...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to Vicia sativa ILP molecular marker primers and an application thereof in variety identification. The 41 pairs of molecular marker primers are used for identifying 30 Vicia sativa varieties, can identify a plurality of varieties, and have the advantages of accuracy, high efficiency, rapidness, low cost, conveniencein operation and the like.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an ILP molecular marker primer of pea quiver and its application in variety identification. Background technique [0002] Arrowhead pea (Vicia sativa L.) is a high-quality annual, self-pollinating, diploid leguminous herbage, which has the characteristics of high nutritional content, rich protein content, strong stress resistance and wide planting area. It can be used as Green manure, forage grass and silage, etc., have a very important position in my country's agricultural ecosystem. With the rapid development of animal husbandry, the breeding and approval of new varieties of Arrowpipe pea has entered a new stage, but the methods for the identification of new varieties and the authenticity of varieties are still relatively lagging behind. At present, it is mainly identified from the morphological point of view through the difference of field traits. However, this method...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 刘文献韦兴燚金小煜王彦荣
Owner LANZHOU UNIVERSITY
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